Outcome In Ovarian Cancer Patients Research Articles

Inferior Vena Cava Filter Placement and Risk of Hematogenous Distant Metastasis in Ovarian Cancer

Recent studies have suggested that inferior vena cava (IVC) filter placement in cancer patients is associated with decreased survival time after insertion. Causality, however, is yet to be understood. This study evaluates (i) the patterns of recurrence or progression of disease; and (ii) survival outcomes of ovarian cancer patients who underwent IVC filter placement. A total of 274 patients who underwent primary cytoreductive surgery for epithelial ovarian, fallopian tube, and primary peritoneal cancers were identified for analysis. Anatomic location of the first recurrence or progression of disease, progression-free survival, and overall survival were correlated to IVC filter placement status inserted during the perioperative period. Overall, 38 (13.9%) patients underwent perioperative IVC filter insertion, of which 37 (97.4%) were permanently placed. The most common indication was newly diagnosed venous thromboembolism (VTE) (52.6%). Patients with IVC filter placement for VTE were more likely to develop subsequent deep vein thrombosis (25% vs. 7.2%, odds ratio, 4.31, 95% confidence interval, 1.40-13.3, P = 0.019), have hematogenous distant metastasis as the site of first recurrence or progression of disease (12-mo hematogenous distant metastasis ratio, 45.2% vs. 13.6%, hazard ratio, 5.10, 95% confidence interval, 2.35-11.1, P < 0.001, multivariate analysis), and show decreased survival outcomes (median progression-free survival, 5.7 vs. 15.3 mo, P < 0.001: and median overall survival, 22.1 vs. 47.2 mo, P = 0.002, both multivariate analysis) when compared with patients without IVC filter placement. Our results suggested that IVC filter placement for VTE in the perioperative period of primary cytoreductive surgery for ovarian cancer may be associated with increased risk of hematogenous distant metastasis and resulted in decreased survival.

Coordinated Regulation of β-Tubulin Isotypes and Epithelial-to-Mesenchymal Transition Protein ZEB1 in Breast Cancer Cells

The regulation of β-tubulin isotypes, the primary targets for antimitotic chemotherapeutic drugs like taxanes, has implications for drug response and drug resistance. Over the past 15 years, micro-RNAs have been studied widely as regulators of mRNA levels. For example, the tumor suppressor miR-200c was shown in cell culture to target mesenchymal genes, including ZEB1 [ Cochrane ( 2009 ) Mol. Cancer Ther. 8 ( 5 ), 1055 - 1066 ]. In that work, exogenous miR-200c was also shown to reduce β-tubulin class III, one of its predicted targets. Furthermore, decreased miR-200c and increased β-tubulin class III were associated with poor outcomes for ovarian cancer patients [ Cittelly , D.M. ( 2012 ) Mol. Cancer Ther. 11 ( 12 ), 2556 - 2565 ]. Because miR-200c targets the epithelial-to-mesenchymal inducer ZEB1, we wanted to know whether changes in ZEB1 parallel β-tubulin isotype changes, implicating β-tubulin isotypes in ZEB1-associated cell survival pathways. We found coordinated positive feedback regulation of mRNA for ZEB1 and β-tubulin isotype classes I, III, and IVB in MDA-MB-231 breast cancer cells, commonly used as a model for triple-negative breast cancers. Low levels of paclitaxel (40 nM) were found to significantly reduce mRNA levels for these tubulin genes along with a 2-3-fold increase in miR-200c. ZEB1 silencing also reduced β-tubulin isotype classes I, III, and IVB mRNA, whereas upregulation of ZEB1 was associated with increases in these isotype classes. Our work indicates that paclitaxel-induced reduction of ZEB1 and β-tubulin isotypes are, in part, due to increased activity of miR-200c. These results suggest that in aggressive breast cancers, as modeled by MDA-MB-231 cells, β-tubulin class III is a biomarker for cell survival mediated through ZEB1-induced tumor progression pathways.

Self-reinforcing loop of amphiregulin and Y-box binding protein-1 contributes to poor outcomes in ovarian cancer

The Y-box binding protein-1 (YB-1) transcription factor is associated with unfavorable clinical outcomes. However, the mechanisms underlying this association remain to be fully elucidated. We demonstrate that YB-1 phosphorylation, indicative of YB-1 activation, is a powerful marker of outcomes for ovarian cancer patients. In ovarian cancer, YB-1 phosphorylation is induced by activation of the lysophosphatidic acid (LPA) receptor (LPAR) via SRC-dependent transactivation of the epidermal growth factor receptor (EGFR) that is coupled to MAPK/p90 ribosomal S6 kinase (p90RSK), but not phosphatidylinositol 3-kinase (PI3K)/AKT signaling. Activation of the LPAR/SRC/EGFR/MAPK/p90RSK/YB-1 axis leads to production of the EGFR ligand amphiregulin (AREG). AREG induces ongoing YB-1 phosphorylation as well as YB-1-dependent AREG expression, thus constituting an AREG/YB-1 self-reinforcing loop. Disruption of transactivation of the EGFR and the downstream self-reinforcing loop decreases invasiveness of ovarian cancer cells in vitro and limits ovarian cancer growth in xenograft models. These findings established the regulation and significance of YB-1 phosphorylation, therefore further exploration of this signaling axis as a therapeutic avenue in ovarian cancer is warranted.

Abstract 1421: Interleukin-22, a protective factor for ovarian cancer during TNFa-induced apoptosis.

Abstract Ovarian cancer is the 5th leading cause of death from cancer in women. Overall, only 10% of patients with advanced ovarian cancer are curable; despite the advances in chemotherapy the 5-year survival is still less than 30%. Interleukin-22 (IL-22), a member of the IL-10 family of cytokines is part of the innate immune system and is expressed by the Th17 cells. IL-22 has been shown to regulate several processes such as the acute-phase response, activation of the innate immune system, cell migration, differentiation and gene expression. However, its role in cancer immunology, especially in ovarian cancer has not been studied. The aim of this study is to investigate the expression and the role IL-22 in human ovarian cancer. In our work, we analyzed the expression of IL-22 protein in human ovarian cancer tissues and ascites samples using immunohistochemistry and ELISA; we tested the expression of IL-22 receptor on human ovarian cancer cells via Western Blot, and determined the association between disease outcome and IL-22 gene expression (Affymetrix)in patients with human ovarian cancer. We evaluated the effect of IL-22 on cellular proliferation using MTT assay and we assessed the effect of IL-22 on TNF-a induced apoptosis using caspase 3/7 activity (Caspase Glo) assay and 7AAD staining on multiple human ovarian cancer cell lines. Here we demonstrate that IL-22 was expressed in human ovarian cancer tissue (mostly in the stroma) and that the receptor for IL-22 was expressed on human ovarian cancer cells. We found that high expression of IL-22, as well as the IL-22-receptor in the tumor tissue leads to poor disease outcome in ovarian cancer patients. On the contrary, high expression of IL-22-binding protein (antagonist of IL-22) was associated with better survival in ovarian cancer patients. We showed that treatment with human recombinant (hr) IL-22 protein induced the activation of Stat-3, as well as increased the proliferation of human ovarian cancer cell lines. We also found that treatment with hr IL-22 at 100, 50 or even at 10 ng/mL concentrations inhibited TNF-a induced apoptosis in OVCAR-5 ovarian cancer cell line, and these results were reversed by the addition of IL22 binding protein to the culture. We conclude that interleukin-22 is an important factor in the tumor microenvironment of ovarian cancer; it promotes tumor growth by increasing proliferation and serves as a protective factor for ovarian cancer during TNF-s induced apoptosis. These data suggest that IL-22 is a potential therapeutical target and/or biomarker in human ovarian cancer. Citation Format: Klara Balint, Tamara Seedial, Phyllis Gimotty, George Coukos, Andrea Facciabene. Interleukin-22, a protective factor for ovarian cancer during TNFa-induced apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1421. doi:10.1158/1538-7445.AM2013-1421

Abstract 4843: Genetic variants in Notch signaling pathway and ovarian cancer.

Abstract The Notch signaling pathway drives proliferation, differentiation, apoptosis, cell fate choices, and has a key role in the regulation of stem cells in a variety of cancers. In this study, we applied a pathway-based approach to evaluate the associations between single nucleotide polymorphisms in Notch signaling pathway and ovarian cancer risk, survival, and response to chemotherapy. Towards this, we systematically genotyped 139 SNPs from 10 genes in the pathway in 339 Caucasian ovarian cancer cases and 349 Caucasian healthy controls. Individually, four SNPs were significantly associated with the ovarian cancer risk. The most significant SNP was NOTCH4: rs397081, with an adjusted odds ratio (OR) of 0.38 (95% CI = 0.22-0.66, P=0.00057). A highly significant gene-dosage effect was observed for these top variants (P for trend &amp;lt; 0.00001) with those carrying ≥ 3 risk genotypes having a nearly 6-fold increase in risk (OR = 5.67, 95% CI = 2.58 - 12.44] . Classification and regression tree (CART) analysis further revealed potential higher order gene-gene interactions between these four SNPs, with the highest risk group having almost 7-fold increased risk. Ten SNPs were significantly associated with overall survival, of which the strongest association was identified for NOTCH4: rs3131290 (OR = 0.48, 95% CI = 0.27-0.84). Cumulative effect analysis showed that multiple adverse genotypes had a significant dose-dependent effect on overall survival. The median survival times for individuals carrying ≤3, 4 to 6, and ≥7 of these adverse genotypes were &amp;gt;186.3, 62.82, and 28.58 months, respectively. Similarly, a significant association was identified between a poor response to platinum-based chemotherapy and the number of unfavorable SNPs (P for trend &amp;lt; 0.0001). Our findings suggest that genetic variants in the Notch signaling pathway are associated with ovarian cancer risk and could serve as potential predictors of clinical outcomes in ovarian cancer patients. Citation Format: Enping Xu, Karen H. Lu, Michelle AT Hildebrandt, Maosheng Huang, Xifeng Wu, Dong Liang. Genetic variants in Notch signaling pathway and ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4843. doi:10.1158/1538-7445.AM2013-4843

L11.04 - Ovarian Avatar Models Predictive of Platinum Response in Ovarian Cancer Patients

Ovarian cancer is the most lethal gynecological malignancy. A major contributing factor in the overall mortality of this disease is the failure to identify patient subgroups sensitive to targeted and individualized therapy. While all advanced ovarian, primary peritoneal and fallopian tube cancer patients typically receive platinum/taxane chemotherapy, many will respond suboptimally. Ovarian cancer tissue +/- ascites was collected from patients undergoing primary debulking surgery. Tumor was minced into a slurry and injected +/- ascites intraperitoneally (IP) in 3 SCID mice/patient and were followed for engraftment for up to 1 year. Upon engraftment (founder), tumor tissue was resected, minced and divided for molecular analyses (Affymetrix Human U133 Plus 2.0) and expansion of tumor tissue in additional mice. Mice bearing the expanded xenografts were then treated with weekly paclitaxel (15 mg/kg) and carboplatin (50 mg/kg) for 4 weeks, sacrificed and tumors were harvested. Tumor volume was assessed biweekly during treatment by ultrasound. From March 2010 to December 2012, tumors were implanted from 246 individual patients. After an initial pilot study, the tumor engraftment rate was 74%. Tumor engraftment occurred in varying histological subtypes of ovarian cancer. Tumor engraftment was associated with poor outcome in ovarian cancer patients from which source tumors originated (OR 5.074, 95% CI 1.1-23.3). DNA microarray experiments demonstrated that Avatars clustered in 1 of 3 subgroups defined by the ovarian TCGA dataset: proliferative, dedifferentiated or immunoreactive. Tumors originating from 3 source patients with platinum-sensitive disease, as defined by no evidence of recurrence > 2 yrs, demonstrated responsiveness to paclitaxel/carboplatin chemotherapy with tumor regression. Conversely, tumor from a patient with platinum-resistant disease, as defined by recurrence within 6 months, failed to respond to chemotherapy in vivo. IP-derived, ovarian tumorgraft models may be generated from primary surgical specimens at a high engraftment rate. Failure to engraft had positive prognostic implications for the source patient. Response to platinum-based chemotherapy in ovarian Avatar models appear to reflect the responsiveness of source patients. The ovarian Avatar models may be a useful in identifying subgroups of ovarian cancer patients that would most likely to benefit from targeted and/or novel therapeutics.

Outcomes following percutaneous upper gastrointestinal decompressive tube placement for malignant bowel obstruction in ovarian cancer

ObjectiveThe objective of this study was to evaluate peri-operative and survival outcomes of ovarian cancer patients undergoing percutaneous upper gastrointestinal decompression for malignant bowel obstruction (MBO). MethodsRetrospective chart review was used to identify patients with ovarian, peritoneal, or fallopian tube cancer who underwent palliative decompressive treatment for MBO from 1/2002 to 12/2010. Kaplan–Meier methods were used to estimate the median survival (MS) and multivariate analysis used to determine if any variables were associated with the hazard of death. ResultsFifty-three patients met inclusion criteria. Median length of diagnosis prior to intervention was 21months. Fifteen (28.3%) patients experienced complications and 9 required revision. Forty-nine (92.5%) experienced relief of symptoms after placement, and 91% tolerated some form of oral intake. Following placement, 19 (36%) patients received additional chemotherapy and 21(41%) patients received total parental nutrition (TPN). Thirty-five patients were discharged home/outpatient facility, 16 to hospice care, and 2 died prior to discharge. MS for all patients was 46days. Patients who received chemotherapy had a MS of 169days compared to 33days (p<0.001). We failed to find an association between survival and TPN or performance status. ConclusionsMalignant bowel obstruction is a common complication of ovarian cancer. Management is palliative; risks and benefits of any therapy must be considered. Percutaneous decompressive therapy provides relief from associated symptoms, and allows patients to be discharged home. Median survival in this group is limited, and decisions regarding aggressive therapy should be individualized.

High expression of crystallin αB represents an independent molecular marker for unfavourable ovarian cancer patient outcome and impairs TRAIL‐ and cisplatin‐induced apoptosis in human ovarian cancer cells

Dysregulated apoptotic pathways are regarded as major reasons for chemoresistance development as a particular challenge in ovarian cancer therapy. In search of molecular factors affecting human ovarian cancer cell apoptosis and, consequently, patient survival, we examined tumors of 103 platinum-/taxane-treated ovarian cancer patients by mRNA-array hybridization, qPCR, and immunohistochemistry. We identified high expression of crystallin αB (CRYAB), a proposed negative regulator of tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-mediated apoptosis. By Kaplan Meier analysis, this factor turned out to be significantly associated with poor patient outcome [overall survival (OS) p = 0.001, recurrence-free survival (RFS) p = 0.003]. Elevated hazard ratios (HR) were estimated with regard to OS (HR = 2.11, 95% CI 1.10-4.06) and RFS (HR = 1.92, 95% CI 1.07-3.47) in multivariable analyses. These associations were confirmed in independent, publicly available mRNA data comprising 431 patients for OS (p < 0.001) and 413 for RFS (p < 0.001). Our findings were validated by studying apoptotic events in cultured human ovarian cancer cells which were stably transfected to express elevated CRYAB levels. These data emphasized the crucial role of CRYAB in human ovarian cancer biology since TRAIL- as well as cisplatin-induced apoptosis was significantly impaired as a function of enhanced CRYAB expression. Taken together, we identified CRYAB as an independent biomarker for unfavourable outcome of human ovarian cancer patients. Since TRAIL is currently tested as anti-cancer drug and large proportions of the present patient cohort displayed low CRYAB levels in their tumors, CRYAB may enable the selection of patient subgroups benefiting most from TRAIL-containing therapy.

Prediction Value of XRCC 1 Gene Polymorphism on the Survival of Ovarian Cancer Treated by Adjuvant Chemotherapy

We conducted a prospective study to test the association between three amino acid substitution polymorphismic variants of DNA repair genes, XRCC1 (Arg194Trp), XRCC1(Arg280His) and XRCC1 (Arg399Gln), and clinical outcome of ovarian cancer patients undergoing adjuvant chemotherapy. 195 patients with primary advanced ovarian cancer and treated by adjuvant chemotherapy were included in our study. All were followed-up from Jan. 2007 to Jan. 2012. Genotyping of XRCC1 polymorphisms was conducted by TaqMan Gene Expression assays. The XRCC1 194 Trp/Trp genotype conferred a significant risk of death from ovarian cancer when compared with Arg/Arg (HR=1.56, 95%CI=1.04-3.15). Similarly, those carrying the XRCC1 399 Gln/Gln genotype had a increased risk of death as compared to the XRCC1 399Arg/ Arg genotype with an HR (95% CI) of 1.98 (1.09-3.93). This study is the first to provide evidence that XRCC1 gene polymorphisms would well be useful as surrogate markers of clinical outcome in ovarian cancer cases undergoing adjuvant chemotherapy.

PCN31 Treatments Pattern and Outcomes of Ovarian Cancer Patients in a Patient-Centered Retrospective Research Registry

PCN31 Treatments Pattern and Outcomes of Ovarian Cancer Patients in a Patient-Centered Retrospective Research Registry

Monocytes and the 38kDa-antigen of mycobacterium tuberculosis modulate natural killer cell activity and their cytolysis directed against ovarian cancer cell lines

BackgroundDespite strong efforts to improve clinical outcome of ovarian cancer patients by conventional and targeted immuno-based therapies, the prognosis of advanced ovarian cancer is still poor. Natural killer (NK) cells mediate antibody-dependent cellular cytotoxicity (ADCC), release immunostimulatory cytokines and thus function as potent anti-tumour effector cells. However, tumour cells developed mechanisms to escape from an effective immune response. So highly immunogenic substances, like the 38 kDa-preparation of M. tuberculosis, PstS-1, are explored for their potential to enhance cancer-targeted immune responses. In this study we examined the modulation of different NK cell functions by accessory monocytes and PstS-1. We focussed on NK cell activation as well as natural and antibody-dependent cellular cytotoxicity directed against epidermal-growth-factor-receptor (EGFR)-positive ovarian cancer cell lines.MethodsActivation, cytokine release and cytotoxicity of NK cells stimulated by monocytes and PstS-1 were determined by FACS-analysis, ELISA, Bioplex assay and quantitative polymerase-chain reaction (qPCR). Transwell assays were used to discriminate cell-cell contact-dependent from contact-independent mechanisms. Five ovarian cancer cell lines (A2780, IGROV-1, OVCAR-3, OVCAR-4 and SKOV-3) with different EGFR-expression were used as target cells for natural and antibody-dependent cellular cytotoxicity assays. Cetuximab (anti-EGFR-antibody) was used for ADCC studies.ResultsOur data show that monocytes effectively enhance activation as well natural and antibody-dependent cytolytic activity of NK cells. PstS-1 directly stimulated monocytes and further activated monocyte-NK-co-cultures. However, PstS-1 did not directly influence purified NK cells and did also not affect natural and antibody-dependent cellular cytotoxicity directed against EGFR-positive ovarian cancer cells, even in presence of monocytes. Direct cell-cell contact between NK cells and monocytes was required for NK activation, while released cytokines seemed to play a minor role.ConclusionsOur data suggest that monocytes enhance natural and antibody-dependent cytotoxic activity of NK cells in a cell-cell contact dependent manner. The TLR-agonist PstS-1 provides additional monocyte activation and induces NK activation markers, while NK cytotoxicity remains unaffected. We conclude that monocytes provide accessory function for ADCC exerted by NK during antibody-based cancer immunotherapy directed against EGFR-positive ovarian cancer cells.

971PD - Functional -413 a > T Polymorphism in Hmox1 Gene Is Associated with the Clinical Outcome of Ovarian Cancer Patients Treated with Paclitaxel/Platinum Analogue

ABSTRACT Introduction Heme oxygenase 1 (HMOX1) is a key enzyme involved in the heme metabolism. HMOX1 expression may be induced by oxidative stress and cytokines, and there is increasing evidence that it may exert cytoprotective effects and facilitate survival and growth of some tumor cells. Aim: The aim of our study was to analyze whether clinical outcome of ovarian cancer patients subjected to standard therapeutic procedures may be associated with the functional -413A > T single nucleotide polymorphism (SNP) within the promoter of HMOX1 gene. Methods The study included 135 patients (median age, 54 years; 95% CI, 39-69) with stage I-IV epithelial ovarian cancer who underwent cytoreductive surgery followed by standard paclitaxel/platinum analogue chemotherapy. Patients DNA was routinely isolated from the peripheral blood and -413A > T SNP was genotyped using specific SNP Genotyping Assay (Applied Biosystems). Results The respective frequencies of -413A > T SNP AA, AT and TT genotypes within the patients' group were 34.1% (46/135), 46.7% (63/135) and 19.3% (26/135) and were similar to distribution within the general population control group. Kaplan-Meier analysis has revealed that AA genotype was associated with significantly longer progression free survival time. The median progression free survival time was 22.2 months in AA harboring patients and 14.5 months in patients with AT + TT genotype (p = 0.033 by log-rank test). AA genotype was also significantly associated with longer overall survival time. Three-year overall survival in AA and AT + TT group was 77.9% and 52.5%, respectively (p = 0.014). Conclusions The results of our study show for the first time that response to the first line treatment and clinical outcome of ovarian cancer patients with standard paclitaxel/platinum analogue chemotherapy might be associated with functional -413A > T SNP in the HMOX1 gene. This finding may be of practical importance for prognostication and may suggest that HMOX-1 is a potential target for a supportive clinical intervention. Disclosure All authors have declared no conflicts of interest.

Photoimmunotherapy and Irradiance Modulation Reduce Chemotherapy Cycles and Toxicity in a Murine Model for Ovarian Carcinomatosis: Perspective and Results

Significant toxicities from multiple cycles of chemotherapy often cause delays or early termination of treatment, leading to poor outcomes in ovarian cancer patients. Complementary modalities that potentiate the efficacy of traditional agents with fewer cycles and less toxicity are needed. Photodynamic therapy is a mechanistically-distinct modality that synergizes with chemo and biologic agents. A combination regimen with a clinically relevant chemotherapy cocktail (cisplatin + paclitaxel) and anti-EGFR targeted photoimmunotherapy (PIT) is evaluated in a murine model for ovarian carcinomatosis. Mice received either 1 or 2 chemotherapy cycles followed by PIT with a chlorine6-Erbitux photoimmunoconjugate and 25 J/cm2 light. PIT + 1 cycle of chemotherapy significantly reduced tumor burden, comparable to multiple chemotherapy cycles. Relative to 1 cycle of chemotherapy, the addition of PIT did not cause significant mouse weight loss, whereas 2 cycles of chemotherapy led to a significant reduction in weight. Irradiance-dependence on PIT efficacy was a function of the conjugation chemistry, providing an additional variable for optimization of PIT outcome.

88. Psychological resilience is associated with markers of inflammation and angiogenesis in pre-surgical ovarian cancer patients

Although psychological distress has been linked to biological processes that may promote ovarian cancer, little is known about the role that psychological resilience plays in this context. To address this issue, we examined how scores on a multi-faceted index of psychological resilience related to biomarkers of inflammation (interleukin-6; IL-6) and angiogenesis (vascular endothelial growth factor; VEGF) in a sample of 200 ovarian cancer patients assessed prior to diagnostic surgery. Using confirmatory factor analysis, a psychological resilience factor with the best fit was determined (Chi-square = 7.28, p = .20; RMSEA = 0.05; CFI = 0.99; TLI = 0.98). Factors included in the measurement model as indicators of resilience include psychological well-being (Psychological Well-Being Scales), positive affect (CESD-D positive affect subscale), and the spiritual, emotional, and functional well-being subscales of the Functional Assessment of Cancer Therapy scale. This psychological resilience index was then used in a structural equation model with IL-6 and VEGF regressed on the latent variable, controlling for cancer stage and grade, age, BMI, and fatigue. Greater psychological resilience was significantly associated with lower levels of IL-6 (beta = −0.20, SE=.09, p = 0.026) but was unrelated to levels of VEGF (beta = −0.11, SE=.09, p = .241). This suggests that protective psychological factors may be linked to inflammatory biology in ovarian cancer. Additional research is needed to examine how psychological risk and resilience factors interact to predict clinical outcomes in ovarian cancer patients.

Correlation of pretreatment drug induced apoptosis in ovarian cancer cells with patient survival and clinical response

BackgroundThis study was performed to determine if a chemotherapy-induced apoptosis assay (MiCK) could predict the best therapy for patients with ovarian cancer.MethodsA prospective, multi-institutional and blinded trial of the assay was conducted in 104 evaluable ovarian cancer patients treated with chemotherapy. The MiCK assay was performed prior to therapy, but treating physicians were not told of the results and selected treatment only on clinical criteria. Outcomes (response, time to relapse, and survival) were compared to the drug-induced apoptosis observed in the assay.ResultsOverall survival in primary therapy, chemotherapy naïve patients with Stage III or IV disease was longer if patients received a chemotherapy which was best in the MiCK assay, compared to shorter survival in patients who received a chemotherapy that was not the best. (p < 0.01, hazard ratio HR 0.23). Multivariate model risk ratio showed use of the best chemotherapy in the MiCK assay was the strongest predictor of overall survival (p < 0.01) in stage III or IV patients. Standard therapy with carboplatin plus paclitaxel (C + P) was not the best chemotherapy in the MiCK assay in 44% of patients. If patients received C + P and it was the best chemotherapy in the MiCK assay, they had longer survival than those patients receiving C + P when it was not the best chemotherapy in the assay (p = 0.03). Relapse-free interval in primary therapy patients was longer if patients received the best chemotherapy from the MiCK assay (p = 0.03, HR 0.52). Response rates (CR + PR) were higher if physicians used an active chemotherapy based on the MiCK assay (p = 0.03).ConclusionThe MiCK assay can predict the chemotherapy associated with better outcomes in ovarian cancer patients. This study quantifies outcome benefits on which a prospective randomized trial can be developed.

Abstract 3276: A novel in vivo xenograft mouse model of human high-grade serous ovarian cancer, with clinical, molecular and functional annotation relevant for pre-clinical analysis

Abstract Background: The five year survival rate for women with ovarian cancer (OC) is less than 40%. Crucial to improving the outcomes for ovarian cancer patients is the development of accurate pre-clinical models of human epithelial OC, which can be used to unravel the mechanisms underlying its evolution and behaviour. Methods: We have generated a novel xenograft model of human high-grade serous OC (HG-SOC) in order to enable preclinical molecular and drug response assessment of individual HG-SOC: this includes a comparison of standard OC xeno-transplantion approaches (intra-peritoneal, subcutaneous and sub renal capsular), with the novel use of the rodent ovarian bursa, the orthotopic site. Utilising the rodent bursa may better replicate the relevant microenvironment and increase the success of EOC xeno-transplantion. Other aspects designed to optimize the model include the use of NOD-SCID-IL-2rg recipient mice, systemic estrogen supplementation and transplantation of fresh human HG-SOC fragments which have had no prior in vitro culture. Histological, functional and molecular analysis of the novel xenograft cohort (at baseline and following xenotransplantation) includes histological review; documentation of in vitro Homologous Recombination (HR) DNA repair and drug response capabilities (using novel α-irradiation and explant drug assays); classification according to molecular subtype (Tothill classfier); documentation of NHEJ pathway status, BRCA1/2 status and other DNA repair gene status. In vivo drug treatment studies are being performed, with the choice of treatment targeted to the specific molecular characteristics of the HG-SOC in question. Results: Fourteen consecutive chemotherapy naive potentially HG-SOC samples have been collected. Corresponding clinical data has been collected. Data concerning DNA repair capability and response to DNA damaging agents will be presented, including IHC for markers of DNA damage (γH2AX), DNA repair (RAD51) and apoptosis (capsase 3 cleavage). HG-SOC in this cohort have been classified according to Tothilll (Tothill et al 2008). Eight appropriate HG-SOC have been transplanted and 5 of the first 6 have successfully xenografted, with phenotyping of xenografts underway. In vivo analysis of response to cisplatin treatment and other relevant therapeutics will be presented. Conclusions: A novel xenograft model has been developed of human HG-SOC, which includes comparative characterization of important prognostic features both in the baseline panel of fresh human HG-SOC and in subsequent xenografts. This clinically, functionally and molecularly annotated consecutive xenograft cohort of HG-SOC will provide outstanding utility for the development of improved therapeutic approaches. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3276. doi:1538-7445.AM2012-3276

A DNA Repair Pathway–Focused Score for Prediction of Outcomes in Ovarian Cancer Treated With Platinum-Based Chemotherapy

BackgroundNew tools are needed to predict outcomes of ovarian cancer patients treated with platinum-based chemotherapy. We hypothesized that a molecular score based on expression of genes that are involved in platinum-induced DNA damage repair could provide such prognostic information.MethodsGene expression data was extracted from The Cancer Genome Atlas (TCGA) database for 151 DNA repair genes from tumors of serous ovarian cystadenocarcinoma patients (n = 511). A molecular score was generated based on the expression of 23 genes involved in platinum-induced DNA damage repair pathways. Patients were divided into low (scores 0–10) and high (scores 11–20) score groups, and overall survival (OS) was analyzed by Kaplan–Meier method. Results were validated in two gene expression microarray datasets. Association of the score with OS was compared with known clinical factors (age, stage, grade, and extent of surgical debulking) using univariate and multivariable Cox proportional hazards models. Score performance was evaluated by receiver operating characteristic (ROC) curve analysis. Correlations between the score and likelihood of complete response, recurrence-free survival, and progression-free survival were assessed. Statistical tests were two-sided.ResultsImproved survival was associated with being in the high-scoring group (high vs low scores: 5-year OS, 40% vs 17%, P < .001), and results were reproduced in the validation datasets (P < .05). The score was the only pretreatment factor that showed a statistically significant association with OS (high vs low scores, hazard ratio of death = 0.40, 95% confidence interval = 0.32 to 0.66, P < .001). ROC curves indicated that the score outperformed the known clinical factors (score in a validation dataset vs clinical factors, area under the curve = 0.65 vs 0.52). The score positively correlated with complete response rate, recurrence-free survival, and progression-free survival (Pearson correlation coefficient [r2] = 0.60, 0.84, and 0.80, respectively; P < .001 for all).ConclusionThe DNA repair pathway–focused score can be used to predict outcomes and response to platinum therapy in ovarian cancer patients.

Outcomes of ovarian cancer patients after bevacizumab and chemotherapy in primary vs. recurrent setting – A multi-institutional study

Outcomes of ovarian cancer patients after bevacizumab and chemotherapy in primary vs. recurrent setting – A multi-institutional study

Loss of secreted frizzled-related protein 4 correlates with an aggressive phenotype and predicts poor outcome in ovarian cancer patients.

BackgroundActivation of the Wnt signaling pathway is implicated in aberrant cellular proliferation in various cancers. In 40% of endometrioid ovarian cancers, constitutive activation of the pathway is due to oncogenic mutations in β-catenin or other inactivating mutations in key negative regulators. Secreted frizzled-related protein 4 (SFRP4) has been proposed to have inhibitory activity through binding and sequestering Wnt ligands.Methodology/Principal FindingsWe performed RT-qPCR and Western-blotting in primary cultures and ovarian cell lines for SFRP4 and its key downstream regulators activated β-catenin, β-catenin and GSK3β. SFRP4 was then examined by immunohistochemistry in a cohort of 721 patients and due to its proposed secretory function, in plasma, presenting the first ELISA for SFRP4. SFRP4 was most highly expressed in tubal epithelium and decreased with malignant transformation, both on RNA and on protein level, where it was even more profound in the membrane fraction (p<0.0001). SFRP4 was expressed on the protein level in all histotypes of ovarian cancer but was decreased from borderline tumors to cancers and with loss of cellular differentiation. Loss of membrane expression was an independent predictor of poor survival in ovarian cancer patients (p = 0.02 unadjusted; p = 0.089 adjusted), which increased the risk of a patient to die from this disease by the factor 1.8.Conclusions/SignificanceOur results support a role for SFRP4 as a tumor suppressor gene in ovarian cancers via inhibition of the Wnt signaling pathway. This has not only predictive implications but could also facilitate a therapeutic role using epigenetic targets.

Proteomic analysis reflects different histologic subtypes of epithelial ovarian cancer

Ovarian carcinoma is a significant cause of cancer mortality in women worldwide. As a heterogeneous disease, distinct clinical and molecular characteristics exist among different histologic subtypes. With the developments in proteomics, surfaced-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is sensitive enough to detect minute quantity of proteins from serum or microdissected cryostat sections. Herein we hypothesized that differentially expressed protein profiles exist in ovarian carcinomas with distinct histologic subtypes. Compared with endometrioid carcinoma, two peaks were significantly higher in serous carcinoma with the m/z of 3622Da and 4778Da, reinforcing the need to treat different histologic subtypes of ovarian cancer as different disease entities. Better understanding of these potential diagnostic and therapeutic biomarkers followed with proof-of-target effect will finally contribute to rational combinations of novel therapy and improve individual ovarian cancer patient outcome.